Biophotonics & Bioimaging

Research Projects

Biophotonic Probes: Synthesis and Applications in Biological Imaging

Organic Chemistry and Biophotonics Research at NJIT

We are developing novel organic nonlinear optical probes and bioconjugates that can be used to detect events inside biological systems and for deep tissue in vivo imaging via two-photon fluorescence microscopy. These nonlinear optical probes, i.e. organic two-photon absorbing probes, are more sensitive and more stable than existing detection methods and are able to identify lesser-known activities inside biological systems (cells or tissues). With sponsorship from the Institute of Biomedical Imaging and Bioengineering of the National Institute of Health, we developed probes for two-photon based deep tissue tumor and angiogenesis imaging.


Two-photon Fluorescence Wound Healing Tissue Imaging to 1.6mm Depth

Deep Vascular Imaging

 

PI/Collaborators: 

Dr. Kevin D. Belfield
Dean, College of Science and Liberal Arts
belfield@njit.edu

Dr. Yuanwei Zhang
Assistant Professor, Chemistry and Environmental Science
yuanwei.zhang@njit.edu

Selected Publications

  1. Yue, X.; Armijo, Z.; King, K.; Bondar, M. V.; Morales, A. R.; Frazer, A.; Mikhailov, I. A.; Przhonska, O. V.; Belfield, K. D. “Steady-state and femtosecond transient absorption spectroscopy of new two-photon absorbing fluorene-containing quinolizinium cation membrane probes” ACS Applied Materials and Interfaces 2015, 7, 2833-2846.

  2. Sui, B.; Yue, X.; Tichy, M. G.; Liu, T.; Belfield, K. D. “Improved Synthesis of the Triazacryptand (TAC) and its Application in the Construction of a Fluorescent TAC-BODIPY Conjugate for K+ Sensing in Live Cells” European Journal of Organic Chemistry 2015, 1189-1192.

  3. Sui, B.; Tang, S.; Liu, T.; Kim, B.; Belfield, K. D. “Novel BODIPY-based Fluorescence Turn-on Sensor for Fe3+ and its Bioimaging Application in Living Cells” ACS Applied Materials and Interfaces 2014, 6, 18408-18412.

  4. Woodward, A. W.; Frazer, A.; Morales, A. M.; Yu, J.; Moore, A. F.; Campiglia, A. D.; Jucov, E. V.; Timofeeva, T. V.; Belfield, K. D. “Two-photon sensitized visible and near-IR luminescence of lanthanide complexes using a fluorene-based donor-π-acceptor diketonate” Dalton Transactions 2014, 43, 16626-16639.

  5. Zhang, Y.; Yue, X.; Kim, B.; Yao, S.; Belfield, K. D. “Deoxyribonucleoside Modified Squaraines as Near-IR Viscosity Sensors” Chemistry – A European Journal 2014, 20, 7249-7253.

  6. Frazer, A.; Morales, A. R.; Woodward, A. W.; Tongwa, P.; Timofeeva, T.; Belfield, K. D. “Luminescent Fluorene-Based Bis-Pyrazolyl Aniline Ligand for Aluminum Detection” Journal of Fluorescence 2014, 24, 239-250.

  7. Zhang, Y.; Yue, X.; Kim, B.; Yao, S.; Bondar, M. V.; Belfield, K. D. “Sensing and Labeling Bovine Serum Albumin (BSA) with Fluorogenic Near-IR-emitting Squarine Dyes” ACS Applied Materials and Interfaces 2013, 5, 8710-8717.

  8. Belfield, K. D.; Bondar, M. V.; Haniff, H. S.; Mikhailov, I. A.; Luchita, G.; Przhonska, O. V. “Superfluorescent Squaraine with Efficient Two-photon Absorption and High Photostability” ChemPhysChem, 2013, 14, 3532-3542.

  9. Zhang, X.; Xiao, Y.; Qi, J.; Qu, J.; Kim, B.; Yue, X.; Belfield, K. D. “Long-wavelength, Photostable, Two-photon Excitable BODIPY Fluorophores Readily Modifiable for Molecular Probes” Journal of Organic Chemistry, 2013, 78, 9153-9160.

  10. Yanez, C.O.; Morales, A. R.; Yue, X.; Urakami, T.; Komatsu, M.; Jarvinen, T. A. H.; Belfield, K. D. “Deep Vascular Imaging in Wounds by Two-Photon Fluorescence Microscopy” PLoS ONE 2013, 8(7), e67559. doi:10.1371/journal.pone.0067559

  11. Belfield, K. D.; Bondar, M. V.; Morales, A. R.; Frazer, A. F.; Mikhailov, I.; Przhonska, O. V. “Photophysical Properties and Ultrafast Excited State Dynamics of a New Two-Photon Absorbing Thiopyranyl Probe” Journal of Physical Chemistry C 2013, 117, 11941-11952.

  12. Sui, B.; Kim, B.; Zhang, Y.; Frazer, A.; Belfield, K. D. “Highly Selective Fluorescence Turn-On Sensor for Fluoride Detection” ACS Applied Materials and Interfaces 2013, 5, 2290-2293.

  13. Yue, X.; Yanez, C. O.; Yao, S.; Belfield, K. D. “Selective Cell Death by Photochemically Induced pH Imbalance in Cancer Cells” Journal of the American Chemical Society 2013, 135, 2112-2115.

  14. Zhao, K.; Liu, T.; Wang, G.; Chang, X.; Xue, D.; Belfield, K. D.; Fang, Y. “A Butterfly-Shaped Pyrene Derivative of Cholesterol and its uses as a Fluorescent Probe” Journal of Physical Chemistry B, 2013, 117, 5659-5667.

  15. Morales, A. R.; Yanez, C. O.; Zhang, Y.; Wang, X.; Biswas, T.; Komatsu, M.; Belfield, K. D. “Small Molecule Fluorophore and Copolymer RGD Peptide Conjugates for Ex Vivo Two-Photon Fluorescence Tumor Vasculature Imaging” Biomaterials 2012, 33, 8477-8485.

  16. Ahn, H.Y.; Yao, S.; Wang. X.; Belfield, K. D. “Near-Infrared Emitting Squaraine Dyes with High 2PA Cross Sections for Multiphoton Fluorescence Imaging” ACS Applied Materials and Interfaces 2012, 4, 2847-2854.

  17. Belfield, K. D.; Bondar, M. V.; Morales, A. R.; Yue, X.; Luchita, G.; Przhonska, O. V. “Transient Excited-state Absorption and Gain Spectroscopy of a Two-Photon Absorbing Probe with Efficient Superfluorescent Properties” Journal of Physical Chemistry C 2012, 116, 11261-11271.

  18. Yao, S.; Belfield, K. D. “Two-photon Fluorescent Probes for Bioimaging” European Journal of Organic Chemistry 2012, 3199-3217.

  19. Yao, S.; Belfield, K. D. “Two-photon Fluorescent Probes for Bioimaging” European Journal of Organic Chemistry 2012, in press (invited microreview).

  20. Ahn, H.-Y.; Fairfull-Smith, K. E.; Morrow, B. J.; Lussini, V.; Kim, B.; Bondar, M. V.; Bottle, S. E.; Belfield, K. D. “Two-photon Fluorescence Microscopy Imaging of Cellular Oxidative Stress using Profluorescent Nitroxides” Journal of the American Chemical Society 2012, 134, 4721-4730.

  21. Belfield, K. D.; Bondar, M. V.; Morales, A. R.; Padhila, L. A.; Przhonska, O. V.; Wang, X. “Two-photon STED Spectral Determination for a New V-shaped Organic Fluorescent Probe with Efficient Two-photon Absorption” ChemPhysChem 2011, 12, 2755-2762.

  22. Andrade, C. D.; Yanez, C. O.; Ahn, H.-Y.; Urakami, T.; Bondar, M. V.; Komatsu, M.; Belfield, K. D. “Two-photon Fluorescence Vascular Bioimaging with New Bioconjugate Probes Selective Towards the Vascular Endothelial Growth Factor Receptor 2” Bioconjugate Chemistry 2011, 22, 2060-2071.

  23. Wang, X.; Morales, A. R.; Urakami, T.; Zhang, L.; Bondar, M. V.; Komatsu, M.; Belfield, K. D. “Folate Receptor-targeted Aggregation-enhanced Near-IR Emitting Silica Nanoprobe for One-photon in vivo and Two-photon ex vivo Fluorescence Bioimaging” Bioconjugate Chemistry 2011, 22, 1438-1450.

  24. Biswas, S.; Wang, X.; Morales, A. R.; Ahn, H.-Y.; Belfield, K. D. “Integrin-targeting Block Copolymer Probes for Two-photon Fluorescence Bioimaging” Biomacromolecules 2011, 12, 441-449.

  25. Belfield, K. D.; Bondar, M. V.; Hernandez, F. E.; Przhonska, O. V.; Wang, X.; Yao, S. “Superfluorescent Fluorenyl Probe with Efficient Two-photon Absorption” Physical Chemistry Chemical Physics 2011, 13, 4303-4310.

  26. Andrade, C. D.; Yanez, C. O.; Qaddoura, M. A.; Wang, X.; Arnett, C. L.; Coombs, S. A.; Yu, J.; Bassiouni, R.; Bondar, M. V.; Belfield, K. D. “Two-Photon Fluorescence Lysosomal Bioimaging with a Micelle-Encapsulated Fluorescent Probe” Journal of Fluorescence 2011, 21, 1223-1230. DOI: 10.1007/s10895-010-0801-3

  27. Nguyen, D. M.; Wang, X.; Ahn, H.-Y.; Rodriguez, L.; Bondar, M. V.; Belfield, K. D. “Novel Hydrophilic Bis(1,2,3-triazolyl)fluorenyl Probe for in vitro Zinc Ion Sensing” ACS Applied Materials and Interfaces 2010, 2, 2978-2981. DOI: 10.1021/am100719v

  28. Parthasarathy, A.; Ahn, H.-Y.; Belfield, K. D.; Schanze, K. S. “Two-photon Excited Florescence of a Conjugated Polyelectrolyte and its Application in Cell Imaging” ACS Applied Materials and Interfaces 2010, 2, 2744-2748. DOI: 10.1021/am100784m

  29. Belfield, K. D.; Andrade, C. D.; Yanez, C. O.; Bondar, M. V.; Hernandez, F. E.; Przhonska, O. V. “New Two-Photon Absorbing Probe with Efficient Superfluorescent Properties” Journal of Physical Chemistry B 2010, 114, 14087-14095. DOI: 10.1021/jp107343k

  30. Wang, X.; Nguyen, D. M.; Yanez, C. O.; Rodriguez, L.; Ahn, H.-Y.; Bondar, M. V.; Belfield, K. D. “High Fidelity Hydrophilic Probe for Two-photon Fluorescence Lysosomal Imaging” Journal of the American Chemical Society 2010, 132, 12237-12239. DOI: 10.1021/ja1057423

  31. Wang, X.; Yao, S.; Ahn, H.-Y.; Zhang, Y.; Bondar, M. V.; Torres, J. A.; Belfield, K. D. “Folate Receptor Targeting Silica Nanoparticle Probe for Two-photon Fluorescence Bioimaging” Biomedical Optics Express 2010, 1, 453-462 (feature article).

  32. Belfield, K. D.; Bondar, M. V.; Frazer, A.; Morales, A. R.; Kachkovsky, O. D.; Mikhailov, I. A.; Masunov, A. E.; Przhonska, O. V. “Fluorene-based Metal Ion Sensing Probe with High Sensitivity to Zn2+ and Efficient Two-photon Absorption” Journal of Physical Chemistry B 2010, 114, 9313-9321.

  33. Yao, S.; Ahn, H.-Y.; Wang, X.; Fu, J.; Van Stryland, E. W.; Hagan, D. J.; Belfield, K. D. “Donor-Acceptor-Donor Fluorene Derivatives for Two-Photon Fluorescence Lysosomal Imaging” Journal of Organic Chemistry 2010, 75, 3965-3974 (feature article). DOI: 10.1021/jo100554j

  34. Andrade, C. D.; Yanez, C. O.; Rodriguez, L.; Belfield, K. D. “A Series of Fluorene-based Two-photon Absorbing Molecules: Synthesis, Linear and Nonlinear Characterization, and Bioimaging” Journal of Organic Chemistry 2010, 75, 3975-3982. DOI: 10.1021/jo1005075

  35. Nguyen, D. M.; Frazer, A., Rodriguez, L.; Belfield, K. D. “Selective fluorescence sensing of zinc and mercury ions with hydrophilic 1,2,3-triazolyl fluorene probes” Chemistry of Materials 2010, 22, 3472-3481. DOI: 10.1021/cm100599g

  36. Morales, A. R.; Luchita, G.; Yanez, C. O.; Bondar, M. V.; Przhonska, O. V.; Belfield, K. D. “Linear and Nonlinear Photophysics and Bioimaging of an Integrin-targeting Water-soluble Fluorenyl Probe” Organic and Biomolecular Chemistry 2010, 8, 2600-2608. DOI: 10.1039/paperno

  37. Morales, A. R.; Yanez, C. O.; Schafer-Hales, K. J.; Marcus, A. I.; Belfield, K. D. “Biomolecule Labeling and Imaging with a New Fluorenyl Two-Photon Fluorescent Probe” Bioconjugate Chemistry 2009, 20, 1992-2000. DOI: 10.1021/bc900302t

  38. Belfield, K. D.; Bondar, M. V.; Yanez, C. O.; Hernandez, F. E.; Przhonska, O. V. “One- and Two-photon Stimulated Emission Depletion of a Sulfonyl-containing Fluorene Derivative” Journal of Physical Chemistry B 2009, 113, 7101-7106. http://pubs.acs.org/doi/full/10.1021/jp902060m

  39. Morales, A. M.; Schafer-Hales, K. J.; Yanez, C. O.; Bondar, M. V.; Przhonska, O. V.; Markus, A. I.; Belfield, K. D. “Excited State Intramolecular Proton Transfer and Photophysics of a New Fluorenyl Two-Photon Fluorescent Probe” ChemPhysChem 2009, 10, 2073-2081. http://dx.doi.org/10.1002/cphc.200900032

  40. Morales, A. R.; Schafer-Hales, K. J., Marcus, A. I.; Belfield, K. D. “Amine Reactive Fluorene Probes: Synthesis, Optical Characterization, Bioconjugation, and Two-Photon Fluorescence Imaging” Bioconjugate Chemistry 2008, 19, 2559-2567. http://dx.doi.org/10.1021/bc800415t

  41. Yao, S.; Schafer-Hales, K. J.; Belfield, K. D. “A New Water-Soluble Near-Neutral Ratiometric Fluorescent pH Indicator” Organic Letters 2007, 9, 5645-5648. http://dx.doi.org/10.1021/ol7026366

  42. Morales, A.R.; Belfield, K. D.; Hales, J. M.; Van Stryland, E. W.; Hagan, D. J. "Synthesis of two-photon absorbing unsymmetrical fluorenyl-based chromophores". Chemistry of Materials, 2006, 18, 4972-4980.

  43. Yao, S.; Schafer-Hales, K. J.; Cohanoschi, I.; Hernandez F. E.; Belfield, K. D. "A water-soluble diaminostilbene derivative as a two-photon fluorescent probe". Synlett, 2006, (12), 1863-1866.

  44. Schafer, K. J.; Belfield, K. D.; Yao. S.; Frederiksen, P. K.; Hales. J. M.; Kolattukudy, P. E. "Fluorene-based fluorescent probes with high two-photon action cross-sections for biological multiphoton imaging applications". Journal of Biomedical Optics, 2005, 10, 051402-1-051402-8.

  45. Yao, S.; Belfield, K. D. "Synthesis of Two-Photon Absorbing Unsymmetrical Branched Chromophores through Direct Tris(bromomethylation) of Fluorene", Journal of Organic Chemistry, 2005, 70, 5126-5132.

  46. Belfield, K. D.; Morales, A. R.; Kang, B.-S.; Hales, J. M.; Hagan, D. J.; Van Stryland, E. W. Chapela, V. M.; Percino, J. "Synthesis, characterization and optical properties of new two-photon absorbing fluorene derivatives". Chemistry of Materials, 2004, 16, 4634-4641.

  47. Belfield, K. D.; Schafer, K. J.; Mourad, W.; Reinhardt, B. A. "Synthesis of New Two-Photon Absorbing Fluorene Derivatives via Cu-Mediated Ullmann Condensations". Journal of Organic Chemistry, 2000, 65, 4475-4481

  48. Belfield, K. D.; Schafer, K. J.; Hagan, D. J.; Van Stryland, E. W., Negres, R. A. "New Two-Photon Absorbing Fluorene Derivatives: Synthesis and Nonlinear Optical Characterization". Organic Letters, 1999, 1, 1575.